Biopharmaceutical process apparatuses assembled into a column

ABSTRACT

A bio factory apparatus capable of Up-Stream production of biologic material products or biologics products in a liquid volume and processing of biologic or biologic material products Down-Stream, comprising at least one first Up-Stream producing capsule and at least one second Down-Stream processing capsule, said capsules capable of being stacked in any order and any number within a column.

TECHNICAL FIELD

The present invention relates to a cultivation apparatus for production of a biologic product, a purification apparatus for purifying a biological product and a bio factory apparatus assembly for producing a purified biologic product. The Up-Stream production is performed by micro organism kept productive in a bioreactor after which the produced crude biologic product is forwarded to Down-Stream processing with various separation methods by numerous technical devices. The invention allows for multiple steps of the production and/or processes to be place in one and same flexible manufacturing apparatus operating in at least one column.

BACKGROUND OF THE INVENTION

EP 317874A2 describes a method for continuous production and removal of a biological substance. This is depending on externally separation devices arranged outside the hollow-fibre bioreactor and not integrated within the bioreactor column. There are no pumps integrated into the producing apparatus.

WO2007/039600 discloses an Up-Stream cell culturing device, which integrates maximum one circulation centrifugal pump operating in a non pressurized vessel. The device has no integration of any Down-Stream devices or second circulation pump.

US 20080008028 disclose a fluid agitating element integrated within a bag or rigid container. The device has no integration with Up-Stream or Down-Stream devices as well as integration in a capsule or said capsule in a column.

U.S. Pat. No. 6,669,844 describe two vertical and opposite oriented rods forcing multiple round filter discs axial together. Said rods insure alignment of multiple filter discs into a module (column) for removal of use in one piece and disposed as a whole without the individual discs having to be separated.

Reference is further made to the Stax™ single-use product platform from the US based company Pall and the Zeta-Plus™ single-use product platform from Cuno/3M. Both platforms depend on stainless steel chassis system for bench-top stacking as well as for floor stacking of capsules on top of each other. Said chassis add considerably axial force to the capsules which insure sufficient gasket contact to the capsules sealing surfaces. No capsule to capsule interlocking devices, straps, latches, locking rings are used or described in the product literature. Gaskets are manufactured from solid rubber. The capsules available are exclusively Down-Stream depth filtration media capsules and single sided end cap/manifold capsules with tri-clamp flanged liquid connections. Operation of /flow distribution in said capsules are exclusively in series with manifold configurations bottom in/top out and/or bottom in/bottom out for connection to externally mounted pumping devices. None of the two platform chassis integrate a pump, a fluid conveyance device, a bioreactor, chromatography or ion-exchange technologies, valve(s), conditioning devices, control devices or the like in a capsule being a part of said column.

Disposable bioreactors were considered a novelty, and perhaps even a passing trend, as recently as mid 90ties. As single-use Up-Stream bioreactors and Down-Stream single-use filters have matured and begun to address most of the present day issues, their market acceptance has rapidly expanded. The development of disposable Up-Stream bioreactors and Down-Stream process equipment is, in many instances, driven by reduction in sterilization and cleaning requirements and validation, improved plant flexibility, reduced clean-room facility costs and faster time to market for the end product. However problems remains the future hurdles to overcome and include the ability to add reliable, accurate sensors and pumping devices so that standards can be generated and the process repeatability readily documented.

Presently none of the traditional technical devices for single-use production system supplied to the Biopharmaceutical Process Plants facilitate the desired integration or modularity. This fact excludes the option of compact integration of both Up-Stream and Down-Stream process equipment, which lack of integration demands expensive and extended facilities.

SUMMARY OF THE INVENTION

Some main aspects of the present invention relates to:

A) A bioreactor apparatus having a top capsule and a bottom capsule, the apparatus comprising a cultivation capsule having a production device for producing a biologic product, and a second upstream capsule, wherein said capsules are stacked and locked in a column, with the proviso that the top capsule and bottom capsule are not pump capsules at the same time.

B) A purification apparatus having a top capsule and a bottom capsule, the apparatus comprising a processing capsule having a purifying device for purifying a biologic product, and a second downstream capsule, wherein said capsules are stacked and locked in a column, with the proviso that the top capsule or bottom capsule are not pump capsules.

C) A bio factory apparatus assembly for producing a purified biologic product comprising a bioreactor apparatus and a purification apparatus in one column having a top capsule and a bottom capsule, the apparatus comprising a cultivation capsule having a production device for producing a biologic product, and a processing capsule having a purifying device for purifying a biologic product, wherein said capsules are stacked and locked in the column, with the proviso that the top capsule and bottom capsule are not pump capsules at the same time

Such apparatus can be pre-sterilized and prepared for single use and an advantage of a pre-sterilized single-use apparatus is the elimination of the preparation steps involved with stainless steel process equipment, such as cleaning, assembly, and disassembly, autoclaving and “clean-in-place” (CIP) or “sterilize-in-place” (SIP) operations, as well as associated labour, validation costs, clean water, waste water and material costs.

A single-use production system, such as any one of the apparatus disclosed above or as described in relation to the figures herein, provides a high degree of flexibility for biotech process design. It allows the practitioner to faster evaluate different configurations before deciding on the final design. Then, should the batch size be modified, or new requirements evolve, the invented single-use biopharmaceutical process plant can be modified more easily to fit with new requirements as to its modularity.

The average biopharmaceutical process plant involves a number of individual process steps typically divided into: 1. Up-Stream and 2. Down-Stream.

Selection of production and processing methods is depending on which type of selected micro organism and biological product to be produced.

Pre Up-Stream Process Steps:

-   -   Preparation of media, serum, nutrient and storage of such     -   Preparation micro organism from a cell bank for inoculation         purposes

1. Up-Stream production is characterised as cultivation in order to produce a biologic product such as proliferation of micro organism or expression by micro organism of a biologic product. Such as pharmaceutical products, such as therapeutic proteins, antibiotics, hormones, enzymes, micro organism or bio mass. Some product of interest may be expressed by micro organisms extra-cellular or intra-cellular (cytoplasmic or periplasmic in the case of bacterial cells). Typically cultivation steps are:

-   -   Inoculation with and proliferation of micro organisms for         seeding purposes in said production bioreactor     -   Mass production of micro organisms for seeding and inoculation         of larger scale production bioreactor     -   Inoculation of said production bioreactor followed by         cultivation of the micro organism in said bioreactor receiving         the desired nutrient

Pre-Down-Stream process step is typically dominated by harvest and separation of contaminants such as un-wanted micro organism, metabolite products and other debris in the crude cultivation broth from the Up-Stream production:

-   -   Crude product recovery (harvest)     -   Separation by filtration     -   Storage of recovered product     -   Buffer preparation and storage of such

2. Down-Stream processing is characterised by a variety of purification methods. In the case of intra-cellular products, the micro organism may be lysed/disrupted and the lysate solution must be clarified to remove the micro organism debris and other contaminants. Depending on the process, additional clarification steps may be needed, such as:

-   -   Pre-filtration/clarification with membrane filters     -   Dilution with buffer (dia-filtration)     -   Intermediate storage     -   Up-concentration of crude product with membranes or centrifuges     -   Membrane absorbers     -   Separation processes     -   Polishing via ion exchange based on chromatography     -   Sterile filtration, removal of impurities with membrane filters     -   Final product storage

Some key biopharmaceutical product groups in biotechnology are:

-   -   Anti-coagulants     -   Blood factors     -   Cytokines     -   Fusion proteins     -   Growth factors     -   Hormones     -   Monoclonal or polyclonal antibodies     -   Polysaccharide vaccines     -   Recombinant vaccines     -   Therapeutic enzymes     -   Therapeutic antibodies     -   Stem cells

Any one of these biologic products can be made by application of the above bioreactor apparatus and/or purification apparatus and/or by combining these technologies into at least one column.

The bioreactor apparatus as well as the purification apparatus and the combined use according to the present invention is particularly well suited for CMOs (Contract Manufacturing Organizations), where a number of different products are produced and the key issues is flexibility, competitive prices and the ability to avoid cross-contamination.

DEFINITIONS RELEVANT FOR THE PRESENT INVENTION

-   -   The term “active liquid pump” as used herein refers to a         centrifugal pump, single membrane pump, dual membrane pump,         peristaltic pump all containing a driving device or force     -   The term “anchorage dependent” as used herein refer to micro         organism with high affinity to anchor, adhered onto a surface,         such as immobilised cells or stem cells or micro organisms in         general further divided into adhering and semi-adherent cell         lines     -   The term “bag” as used herein refers to a flexible (most often         single-use) container made from plastic foil serving various         purposes, media preparation and storage, bioreactor vessel,         product storage     -   The term “batch” as used herein refers to a bioreactor to which         no fresh medium is added and no cultured liquid removed     -   The term “biologics” and “biologics product” as used herein are         used interchangeable and refers to and includes a wide range of         medicinal and therapeutic products and drugs such as:         antibiotic, antibodies, recombinant antibodies, monoclonal         antibodies, vaccines, proteins, recombinant proteins, proteins         molecules, blood components, allergenics, somatic (adult) stem         cells, stem cells in general, tissues created by biological         processes     -   The term “bio mass” as used herein means renewable energy         sources, biological products derived from living, or recently         living organisms, such as wood, waste, (hydrogen) gas, and         alcohol fuels     -   The terms “bioreactor” or “fermenter” as used herein are used         interchangeable and means a physical device, a container, a bag,         a vessel which support biologically active environment and         houses micro organism performing a cultivation process typically         with the micro organism suspended in the media or containing         inside a permeable three-dimensional matrix, skeleton or         scaffolding body     -   The term “buffer” as used herein refers to a liquid containing a         weak acid with resistance to pH changes for stabilising target         molecules, required for a number of filtering and/or         purification steps and adjustment of final product quality     -   The term “capsule” as used herein refers to a series of         containers integrating processing device(s) which may be         circular and cylindrical or non circular or cylindrical, such as         box shaped     -   The term “cell density” describes the cell mass or amount of         cells     -   The term “centrifuge” as used herein means an apparatus using         centrifugal forces to separate substances with very little         difference in density by applying centrifugal force to the         suspension     -   The term “chassis” as used herein means a stainless steel frame         comprising two vertical rod compression based system to         encapsulate capsules, such as the STAX and Cuno capsules     -   The term “chemical compound” as used herein refers to a pure         chemical substance consisting of two or more different chemical         elements, such as organic molecules     -   The term “chromatography” as used herein refers to methods for         purification or capture involving a device that holds         chromatographic active materials, typically one or more steps is         needed in a purification process for protein capture and/or         separations.     -   The term “cross flow mode operation” as used herein refers to         operation principle for a filter unit     -   The term “column” as used herein refers to a container, e.g.         cylindrical container, assembled from two or more capsules         stacked on top of each other into a column, a stack     -   The term “cultivation” and “culturing” as used herein are used         interchangeable and refers to hosting of micro organism in a         bioreactor for production purposes, such as expression or         proliferation     -   The term “dia-filtration” as used herein refers to a method         where water or buffer is added to the biologic product during or         prior to Down-Stream membrane filtration in order to remove         solutes     -   The term “disposable” as used herein refers to a product         manufactured from organic materials preferably taken from the         group of thermo polymers, thermo setting polymers, and elastic         polymers     -   The term “expression” as used herein refers to production of         molecules by micro organism being cultivated, but not to         proliferation of micro organism     -   The term “fed-batch” as used herein refers to a bioreactor to         which fresh medium is added and no cultured liquid removed until         end of the process.     -   The term “fermentation” as used herein refers to large scale         cultivation of micro organisms or single-celled creatures for         industrial purposes in a metabolic process     -   The term “film” as used herein refers to thermoplastic film or         foil made using an extrusion process typically in one or several         layers of different material for different purposes. Films may         be permeable or impermeable and translucent or coloured in         thickness below 1 mm.     -   The term “fluid” as used herein means liquids as well as gases     -   The term “filter” as used herein means a processing device which         by size exclusion separates particles suspended in a fluid in a         method named filtration or separation     -   The term “filtrate” as used herein refers to the part of a         suspension that passes through a filtration process, also called         permeate     -   The term “filtration” as used herein refers to mechanical         separation, size exclusion, fractionating of solids from fluids,         such as liquids or gases, by passing the feed stream through a         porous material such as a porous, fibrous or granular substance,         which retains selected solids and allows other solids and         desired fractions of the fluids to pass through. Membrane filter         products such as micro-filtration, ultra-filtration,         nano-filtration, dia-filtration, gel-filtration are well known         involving operation methods such as dead-end filtration,         cross-flow filtration,     -   The term “fluid conveyance device” as used herein refers to         fluid pumping devices of various principles     -   The term “ion-exchange” as used herein refers to the exchange of         ions between molecules, a solutions and a complex in the form of         an insoluble sorbent, resin with typical physical appearance as         beads or a membrane. The trapping of anions/cations takes place         only with simultaneous releasing of other anions/cations; thus         the process is called ion-exchange.     -   The term “impurities” as used herein refers to any substance         that is not part of the biologic product, such as virus, HCP,         DNA, RNA, Endotoxin     -   The term “macro pore size” as used herein are ranging between         10-500 μm in diameter     -   The terms “media”, “growth media”, and “nutrient” as used herein         are used interchangeable and refers to a sterile complex mixture         containing mostly water, carbon sources, various gases such as         oxygen and additives such as; vitamins, hormones, growth         factors, plant hydrolysates, animal serum, antibiotics,         antioxidants, antifoams, cell stabilizers and other components         for cultivation of micro organisms. Some media are serum based,         some are serum free, animal free, and protein free or chemically         defined     -   The term “meso pore size” are ranging between 1-10 μm in         diameter     -   The term “micro pore size” are ranging below 1 μm in diameter     -   The term “membrane” between refers to a boundary layer, which         serves as a selective barrier and remains impermeable to         specific particles, molecules, or substances when exposed to the         action of a driving force (like a pump). Also a surface layer         shaped as membrane(s) (typically attached to a more rigid         carrier) with selected surface area, pore size, porosity, charge         in order to increase contact and hereby affinity, adsorption,         absorption of selected molecules, particles or the like.     -   The terms “micro organism” or “microbial cell” as used herein         are used interchangeable and is typically divided into: 1.         living single-celled organisms, microbes such as; fungus, algae,         moss, plankton, yeast, protozoa, eukaryotes, archaea, micro         animals, extremophiles and plant cells or the like—2. adherent         or semi adherent or suspended living cells such as animal cells,         insect cells, mammalian cells, human cells, stem cells in         general, embryonic stem cells, induced pluripotent stem cells,         adult (somatic) stem cells in general, regenerative stem cells,         tissue-derived stems cells, stromal vascular fraction stem cells         or the like—3. prokaryotes and a variety of bacteria such as E.         coli or the like—most of the above generically modified to solve         specific tasks and product needs.     -   The term “non-woven” as used herein refers to a porous sheet,         felt or web made from spun-bonded fibres, melt-blown fibres or         other methods with fibre diameter ranging 0.1-50 μm, such as         5-50 μm in diameter, bonded together by chemical, thermal and/or         mechanical methods, which are neither woven nor knitted     -   The term “perfusion mode operation” as used herein refers to the         operation principle for a bioreactor, the media are continuously         exchanged, fresh nutrients added and the crude product is         harvested throughout the cultivation period     -   The term “polishing” as used herein refers to the final         purification step(s) involving an affinity or other refined         chromatography methods     -   The term “porosity” as used herein refers to a measure of the         void spaces in a material expressed in percent 0-100%     -   The term “pre-filtration” as used herein refers to the early         separation of waste (e.g. used and dead micro organisms, damaged         micro organisms) from the crude cultivation product     -   The term “protein” as used herein refers to complex organic         macromolecules often primary ingredient in therapeutic medicine         further proteins are present in each living cell of all         organisms without exceptions and in the cells, the proteins are         abundant in variety or types.     -   The term “passive liquid pump” as used herein refers to a         collection container capable of receiving and despatching a         liquid volume with no active driving device included     -   The term “purification” as used herein means a central part of         the Down-Stream processing that takes a crude cultivation         supernatant or cell homogenate (chaotic slurry of tissue and         cells) and up-concentrates, isolates the biologic product in a         fairly pure form. Involves processes such as filtration,         ion-exchange and chromatography in several difference forms     -   The term “separation” as used herein means dividing fluid borne         particles of different size by membrane filtration or         centrifugal separation based on particle size, mass difference         into at least two separate particle containing fluid streams         further separation by affinity adsorbing the target compound     -   The term “single-use” as used herein refers to a product         designed for use only once and to be disposed after use         typically delivered pre-sterilized and ready to use     -   The term “suspension” as used herein refers to particles, micro         organism suspended in a fluid or mobilised in a fluid     -   The term “suspension dependent” as used herein refers to micro         organism, cell lines suspended in a fluid with no or little         affinity to anchor or adhere to surfaces     -   The term “stem cell(s)” as used herein refers to cells found in         all multi cellular organisms. They are characterized by the         ability to renew themselves through mitotic cell division and         differentiate into a diverse range of specialized cell types.         Stem cells of current international interest originate from         humans and non human primates such as rat, rabbit, bovine, goat,         sheep or marine creatures     -   The term “stacked” as used herein means two or more capsules         positioned close, packed closely next to the neighbouring         capsule or on top of the neighbouring capsule     -   The term “locked” as used herein means two or more capsules         engaged mechanically for operation and/or transportation and/or         storage     -   The term “stacked and locked in a column” as used herein means         two or more capsules which are packed closely and engaged         mechanically and sealed internally with a sealing arrangement,         e.g. gaskets     -   The terms “tri-clamp port, flange” as used herein refers to an         industry method of quick connection of a hose to a conduit, a         container or a pump, the Tri-clamp flanges are separated by a         circular gasket and kept together by a two piece clamp with         internal conical walls.

BRIEF PRESENTATION OF THE FIGS. 1-10

1. Bio factory apparatus stacked and locked in a column with external mounted independent active fluid conveyance pump

2. Bio purification apparatus stacked and locked in a column comprising a selection of capsules for purification purposes, and a capsule comprising a centrifugal pump driven by an electrical motor integrated in separate capsules

3. Bio factory apparatus stacked and locked in a column comprising a first capsule integrating an active membrane pump and a passive membrane operating in an expansion volume in a second capsule

4. Bio factory apparatus stacked and locked in a column comprising a first pump control capsule, in particular the capsule integrates an active membrane pump, and one external mounted fluid collection container

5. Showing a part of the bio factory apparatus, which is a centrifugal pump driven by an electro mechanical motor integrated into separate capsules

6. Showing a part of the bio factory apparatus, which is a first capsule comprising a centrifugal pump driven by a combined magnetic and electrical device housed in a second capsule

7. (a and b) Showing a part of the bio factory apparatus, which is a column illustrating stacking and locking and sealing method

8. Showing a part of the bio factory apparatus, which is an inflatable sealing gasket for use between capsules

9. Showing a part of the bio factory apparatus, which is a set of two identical capsule covers facing towards each other comprising ports for internal integration of cartridges in between

10. Showing a part of the bio factory apparatus, which is a set of two identical mirrored capsule cover parts, which both comprises one or more sets of cylindrical push-in female tube ports

DESCRIPTION OF THE INVENTION

In one aspect the present invention relates to a bioreactor apparatus having a top capsule and a bottom capsule, the apparatus comprising a cultivation capsule having a production device for producing a biologic product, and a second upstream capsule, wherein said capsules are stacked and locked in a column, with the proviso that the top capsule and bottom capsule are not pump capsules at the same time.

Thus, the top capsule may be a cultivation capsule and the bottom capsule may be the second upstream capsule or may be the third or so forth capsule. Alternatively, the cultivation capsule may be located between the top and bottom capsules and may consist of one or more cultivation capsules.

The bioreactor apparatus is preferably for single use, or is pre-sterilized or both. The bioreactor may be any suitable bioreactor, such as a bioreactor selected from a batch, fed-batch, or continuous mode, e.g. continuous perfusion.

The bioreactor may be connected to a purifying apparatus according to the present invention or may be connected to any prior art purifying apparatus such as STAX or Cuno capsules.

In further embodiments of the invention, the bioreactor apparatus performs in continues mode for a limited time (days, weeks or years) the Up-Stream bulk biologic production alone or combined with the Down-Stream processing of the crude biologic products. Up-Stream production is such as:

-   1. hosting of and proliferation of (also genetically modified) micro     organisms for various pharmaceutical purposes such as mass     production of mammalian (including human) cells such as: stem cells,     tissue cells, cartilage cells, skeletal muscle cells intended for     manufacturing of, growth, regeneration of mammalian/human body     parts, organs parts or b; mass production of micro organisms or     living cells as part of a seed train strategy -   2. hosting of and cultivation of micro organisms for expression of     biologics in general, such as therapeutics in general, personalized     medicine, and in specific antibodies, monoclonal antibodies,     vaccines and recombinant products in general, such as antibodies,     DNA, antigens and proteins in general, hormones, enzymes and     vaccines in general and food ingredients and the like -   3. hosting of and proliferation of micro organisms designed for     industrial applications such as production of bio mass in general     such as for energy production purposes

In a further embodiment the production device is in the top capsule.

In another embodiment the production device is in the second or further capsule. Thus, the bioreactor may have just one cultivation capsule or may have several cultivation capsules, such as 2-10 capsules, e.g. 2-8, or 1-4.

In a further embodiment the second up stream capsule is selected from a manifold capsule, cultivation capsule, active fluid conveyance capsule, passive fluid conveyance capsule, fluid collecting container, valve capsule, column end capsule, conditioning capsule, size adaptor capsule, power supply capsules, data acquisition capsules, control capsule and storage capsule.

In a further embodiment the bioreactor apparatus has two or more circular capsules that may be stacked and locked in numbers until 35. Such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25.

In a still further embodiment the bioreactor apparatus has two or more non-circular capsules may be stacked and locked in numbers until 35. Such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25. The method facilitates for square or rectangular capsules at set of four parallel rods to transfer the forces to lock the capsules together between a first fixed base plate at the bottom and a second movable base plate at the upper end of the capsule assembly.

Dimension of circular capsules suitable for desk-top application are ranging 100 to 500 mm and for floor application ranging 200 to 1,000 mm diameter. Dimension of capsules suitable for desktop application is ranging from 150 cm² cross section to 10,000 cm², e.g. from 150 cm² cross section to 1,000 cm² and for floor application ranging from 500 cm² to 50,000 cm², e.g. from 500 cm² to 5,000 cm².

Identical purpose, but different volume capsules offer different processing capacity and cost. Different diameter or cross section format capsules may be interconnected for capacity optimisation or other purpose. An adaptor capsule not having the same dimension on each side will allow two different dimension capsules to interconnect.

The above presented assembling methods promotes disposal of the bio factory apparatus in one complete assembled column. This feature is highly beneficial in specific with micro organism, cultures and processes applying hazards to the work force or facilities in general.

In a further embodiment the bioreactor comprises a further up-stream function selected from cultivation, separation, filtration, conditioning, sparging, diffusion, active fluid conveyance, passive fluid conveyance, collection container, valve, column end, manifold, storage function, heating, cooling, power supply, data acquisition and controlling, wherein the up-stream function or functions are located in one or more capsules.

The production device comprises a porous matrix or a matrix body, such as envelopes containing at least one porous matrix or a matrix body for retaining micro organisms. In a further embodiment the envelopes comprises a three-dimensional porous matrix or scaffolding. In further one embodiment the envelopes contain micro carriers or growth bodies.

According to a further embodiment of the present invention, adherent, semi-adherent, suspension type of micro organisms are kept enclosed at high micro organism density in a three-dimensional matrix with desired pore structure for cultivation said matrix exposed to a media flux, a cultivation method known as perfusion mode. Flux (media velocity per cm² matrix) range from 1 to 1000 cm/min, typically 1 to 100 cm/min, where 5-50 cm/min will support hosting of >1×10⁸ CHO (Chinese Hamster Ovary) cell density and support up to 90% dO₂ inlet concentration and reduction of the dO₂ after passage of the matrix. For other micro organism flux must be even higher such as for algae. Said matrix is encapsulated, integrated inside envelopes in its desired shape inside the invented capsule. The flux and desired shape insures any of and all of the micro organisms are supplied with needed nutrient and kept free from un-wanted results of their activity. By majority adherent and semi-adherent micro organisms will prefer to anchor on the matrix surface and partly micro-suspended within the matrix pores. The suspension type of micro organism will prefer at least partly to be both micro-suspended and agglomerated in the matrix pore volume.

In one preferred embodiment of the invention the production device in the cultivation capsule(s) is a depth filter with porosity ranging 30-99%, such as 50-99%, typically 80-98%, e.g. 80-95%, such as non-woven sheet material. Said fibre based non-woven sheet material having thickness ranging from 1-150 mm preferably ranging 3-50 mm assembled from one or more non-woven layers. Bulk non-woven sheet density ranging from 0.01 g/cm³ to 10 g/cm³. Such non-woven sheet material may have symmetric or asymmetric pore and density properties. Said asymmetrical properties obtained by one or more layers of thinner non-woven sheet material in order to obtain the final desired thickness. The particular production device core design, being integrated into the cultivation capsule(s), is assembled from such as either of:

-   -   Soft matrix bodies based on porous sheet material, such as woven         or non-woven materials, comprising a first fluid inlet and a         second fluid outlet;         -   1. Sets of stacked discs separated by first inlet and second             outlet fluid conveying guides/spacers/cores, said first             inlet fluid guide in fluid contact with supply volume and             said second fluid guide in contact with drainage volume             (reference is made to FIG. 1)         -   2. Straight pleated sheets separated by inlet and outlet             fluid flow guides and shaped into cylindrical bodies         -   3. Curved, wave shape pleated sheets spaced apart by inlet             and outlet fluid flow guides and further shaped into             cylindrical bodies         -   4. Sheets rolled in spiral-wound manner with said sheets             spaced apart by inlet and outlet fluid flow guides     -   Semi rigid matrix bodies based on fibres, particles, beads,         grains, spheres comprising a first fluid inlet and a second         fluid outlet     -   Rigid matrix bodies based on beads, grains, spheres solid         interfaced in the contact points of said beads, grains, spheres         comprising a first fluid inlet and a second fluid outlet     -   Envelopes with thin porous walls comprising a first fluid inlet         and a second fluid outlet encapsulating porous matrix based on         fibres, particulates, beads, grains, spheres, micro carriers

Said porous three-dimensional matrix bodies may be internal surface area coated or treated in order to obtain advantageous features for said micro organism cultivation.

Said production device core comprises dimension of 1-500 mm in height and 10-1,000 mm in width and further shaped such as round or square or rectangular or the like.

Linear scaling up from laboratory scale to “small scale production” bio-reactors allow up-scaling from one non-woven disc diameter 47×4 millimetre thick/6 cm³ to a multi disc set-up diameter 575×1,000 millimetre height/150,000 cm³ creating the impressive scaling number of 1:25,000. With bioreactor volume reduction of 1:40 the invented bio reactor enclosed in the capsule is equivalent (in biologic production capacity) to a suspension tank of 150,000×40=6 m³. The smallest bio-reactor body sized at 0.1 cm³ and the largest being sized at diameter 1,000×2,000 millimetre height being app 1.5 m³ creating the scaling number of 1:15,000,000.

In one preferred embodiment of the invention, the matrix preferably comprises materials selected from the following groups:

-   -   Synthetic polymers in general, such as; resins, polycarbonate,         polypropylene (PP), polyethylene (PE), polyethersulfone,         polyethylene terephtalate/polyester (PET), polyoxyethylene,         polyacrylonitrile, Polysulfone, ethylene vinyl acetate,         cellulose acetate, polytetrafluoroethylene,         Polychlorotrifluoroethylene, Polystyrene, Polycaprolactone         (PCL), Polyfluoroethylenepropylene, polyvinylidene fluoride,         polyvinylidene chloride (PVC), Polyamides, acetal, acrylics and         further thermoplastics in general     -   Synthetic elastomers, rubber in general, such as silicone,         ethylene propylene rubber (EPM), ethylene propylene diene rubber         (EPDM), Fluoroelastomers (Viton), neoprene, polyurethane,         nitrile, butyl,     -   Natural polymers (also bio polymers) in general, any form of         fibres or particles, such as; rubber, latex, cellulose,         cellulose variations, dextran, chitin, collagen, fibrin,         keratin, starch, paper, cotton, wool, flax, hemp, coconut, jute,         resins, viscose rayon, human and animal organic structures such         as bone, tissue and further carbon and carbon element containing         structures in general     -   Synthetic bio degradable polyester (bio plastics) in general,         like; aliphatic polyester such as polylactic acid polymers and         polyhydroxyalkanoates families, polybutylene succinate, etc     -   Human or animal bone or skeleton substitutes such as         hydroxylapatite and/or calcium phosphate and/or carbonated         calcium variants further insects skeleton materials like chitin     -   Ceramics based on powders, grains and/or fibres, such as         non-oxides and/or oxides and/or nitrides based on, SiC,         Titanium, Aluminium, Silicon, Zirconium and their combinations

In general increased micro organism productivity has little effect on cell debris losses from the matrix based bioreactor in the perfusion flow operation mode. The matrix comprise various functionalities, such a insuring attractive surfaces and pore volume, insuring no gradient media distribution though the matrix, insuring even flux all over matrix inlet surface, acting partly as a depth filter. With constant flux, re-circulation media mass flow passing the bioreactor said filtration efficiency increases over time until close to 100% trapping efficiency.

In one preferred embodiment of the invention the production device in the cultivation capsule is a depth filter with porosity ranging 30-99%, such as 50-99%, typically 80-98%, e.g. 80-95%. The matrix porosity insures 80-98% trapping efficiency for 10-30 μm particles, such as the typical micro organisms size, in the first pass depending on velocity. As to the method of perfusion the re-circulation allows the same liquid to pass continuously within selected 1 to 10,000 seconds intervals resulting in further 80-98%, such as 80-95%, trapping efficiency for the remaining suspended particles in the fluid. Within limited re-circulation the trapping efficiency reaches close to 100%. This feature is considered an important part of the invention.

By adding pulses, rapid velocity changes to the liquid (passing the matrix) said matrix is able to overcome the objective depth filter properties and “loose” a desired amount of micro organism, a product or desired particles for various purposes. Such as hosting of and proliferation of (genetically modified) micro organisms for various pharmaceutical purposes such as mass production of mammalian (including human) cells such as: stem cells, tissue cells, cartilage cells, skeletal muscle cells for manufacturing, growth, regeneration of mammalian/human body parts, organs parts or b; mass production of micro organisms or living cells as part of a seed train strategy.

In one preferred embodiment of the invention the bioreactor is supporting cultivation of micro organism depending on certain wave length for photo synthesis. This is supported by integration of translucent capsule materials or light transmitting devices and external light sources or supported by in capsule integrated light sources such as wave length adjustable LED lamps.

The bioreactor apparatus and/or purification apparatus typically comprises a capsule having an active fluid conveyance. In a further embodiment such active fluid conveyance capsule is an active reciprocating fluid conveyance capsule. The active reciprocating fluid conveyance capsule(s) is stacked and locked in any order and any number within the column and comprising liquid nutrient media and micro organisms, the liquid media further corresponding with one or more containers capable of receiving and despatching the liquid media in a such a way that the liquid media is conveyed in reciprocating directions through at least said top capsule and bottom capsule.

In another embodiment such active fluid conveyance capsule is a rotating wheel pump capsule. The rotating wheel pump capsule is stacked and locked in any order and any number within the column and comprising liquid nutrient media and micro organisms, the liquid media further corresponding with one or more containers capable of receiving and despatching the liquid media in a such a way that the liquid media is conveyed in reciprocating directions through at least said top capsule and bottom capsule.

The general fluid movement between containers and filter platform capsules is traditional done by the use of external oriented peristaltic pumps. The peristaltic pump benefit from the easy exchange of the liquid conveying elastomeric material hose(s) through the peristaltic pump is limited severely by the heavy bulky pump frame, motor and hose head.

In a further embodiment the active fluid conveyance comprises stand-alone external oriented pumps connected to the bioreactor apparatus and/or purification apparatus via hoses. In another embodiment the active fluid conveyance comprises capsule integrated devices, such as:

-   -   diaphragm membrane pump     -   centrifugal wheel pump     -   piston pump     -   peristaltic pump

According to further embodiment the diaphragm membrane pump offers excellent low shear stress performance comprising desired integration into a capsule collaborating in a column via conduits with other producing or processing capsules. The diaphragm membrane pump further offers the pulsing effect suitable for:

-   -   re-location of micro organisms inside one capsule helping         different location     -   removal of micro organisms, biologic product, bulk product,         concentrated product from a producing or processing capsule to         the next capsule in the process train

The membrane preferably based on a flexible element manufactured from an elastic material. Said flexible element may take shape such as a flat element, a curved element, or as a cylindrical element prior to expansion.

According to another embodiment (reference is made to FIG. 3) the bioreactor column comprises a first capsule integrating an active membrane pump. A second capsule integrates a larger diameter passive membrane operating as expansion volume. A third capsule integrates a cultivation capsule. A fourth capsule combines manifold & valves devices and a fifth capsule integrates filter for the size exclusion process.

The diaphragm pump operates oscillating based on rear side membrane pressure variation from atmospheric pressure to typically 5 bar pressure. The membrane oscillates controlled by the variation in drive gas pressure on the dry side and the driven fluid on the opposite wet side. The second capsule gas pressure chamber is pre-pressurised (low pressures such as 0.01-1 bar) and will in passive stage force the elastic membrane towards the inner housing part. The pumped fluid conveyed from first capsule is passing the third cultivation capsule (though entirely according to the column design) to the second capsule passive fluid chamber. Hereby the empty gas expansion chamber becomes pressurized according to the liquid volume the active pump is conveying. When the first capsule pump membrane is fully expanded a corresponding liquid volume is stored in the passive pumps wet chamber. As soon as the first capsule membrane becomes drive gas pressure less the stored liquid will return passing the desired capsule or external device controlled by and according to the column design.

The passive membrane is preferably at least 10% larger in diameter than the active membrane in order to facilitate general low pressure operation.

In a further embodiment (reference is made to FIG. 4) the column comprises a first capsule integrating an active membrane pump and one external mounted fluid collection container. The diaphragm pump operates oscillating with rear side membrane pressure variation from atmospheric pressure to typically 5 bar pressure. The membrane oscillates controlled by the variation in drive gas pressure on the dry side and the driven fluid on the opposite wet side. The liquid expansion chamber is not pressurised and positioned with a liquid head (such 0.5-10 meter) above the column. The pumped fluid conveyed from the first capsule pump is passing the second capsule is lifted into the fluid collection container against the force of gravity. Hereby the fluid collection container becomes pressurized according to the liquid volume the active pump is conveying and the distance to the collection volume. When the first capsule pump membrane is fully expanded a corresponding liquid volume is stored in fluid collection containers wet chamber. As soon as the first capsule membrane becomes drive gas pressure less the stored liquid will return passing the desired capsule or external device controlled by and according to the column design.

The above embodiments are important embodiments of the invention as they simplify the invented bio plant apparatus requiring only one active membrane pump, which further eliminate the need for drive gas pressure below atmospheric pressure of said active membrane pump. This simplification as the passive expansion chamber/collection container facilitates the membrane return function of the active pump.

According to another embodiment active fluid conveyance device(s) convey fluids via defined passageways in said capsule. Such as centrifugal pumps with forces applied to a fluid by a rotating wheel (with straight fins or vanes typically for agitation, mixing purposes and curved fins or vanes predominantly for pumping purposes) accelerate the fluid from low to high velocity before being exhausted from the rotating wheel entering the high pressure collecting volume. Said high pressure collection volume chamber is via defined fluid passageways connected to other means such as conduits, flanges or valves. Rotational forces, radial and axial control of the fluid conveying devices(s) performed by methods such as:

-   -   Pump wheel(s) positioned inside a pumping chamber and attached         to a rotating axle penetrating the chamber wall further driven         by motive means outside the pump wheel chamber through a         coupling supported by a rotating bearing, said axle aligned in         bearings inside chamber (reference is made to FIG. 5)     -   Pump wheel(s) positioned inside a pumping chamber and attached         to a rotating axle aligned in between said pump chamber walls         further driven by motive means proximate to, but outside pump         chamber wall and capsule, such as external oriented motive means         for the supply of magnetic forces for levitating and/or rotation         hereby eliminating the otherwise penetrating axle and related         contamination issues (reference is made to FIG. 6)

Increasing the pressure capability of said centrifugal pump the pump wheels are stacked one more in such a manner that the wheels operate in series attached to the same axle via defined passageways in said housing. In one embodiment the capsule contain a two stage centrifugal pump comprising two pumps wheels sharing the same axle—both wheels integrated in the same capsule. Two centrifugal pumps connected in series are able to generate liquid pressures above 3 bar.

According to a further embodiment the capsule integrated fluid conveying device comprises a rotating wheel with vanes in a hollow compartment with inlet and outlet conduits in said capsule. Said integrated fluid conveying device serves multiple purposes including fluid conveyance and further mixing of substances within the same embodiment. Substances for mixing are such as;

-   -   Fluids such as liquids or gases     -   Solids additives, particulate matter, dry powder     -   Solids additives suspended in a fluid     -   Dissolved solid additives constituting a liquid concentrate

into media and/or buffer within the invented column. The active drive and addition of energy to the rotating wheel serves as revolution control, possible temperature increase, stage change from solid to dissolved in liquid. The combined functionality capsule may be coupled to capsule(s) via conduits defining passageways containing one or more valves for potential partial re-circulation of the mixture, for connection with other functions in one or more columns and/or for connection to one or more columns or one or more external storage containers. The combination of active pumping and active mixing actions within the very same capsule eliminates the traditional need for separated and external mixing devices.

Said active fluid conveyance device(s) finds substantial use as single-use pumps integrated into capsules though facilitates usage without being integrated into a column. Reference made to FIG. 6. Such as in a stand-alone application with reference made to FIG. 1.

According to another embodiment the piston pump comprises at least one flat free floating piston or membrane with axial operation controlled by controlling fluid pressure on the side opposite to the liquid or media. Increased controlling fluid pressure reduces the media volume by piston movement. Opposite the decreased controlling fluid pressure increase the volume by the piston movement. The piston movement velocity and variable displacement determined by the controlling fluid. The piston includes a circumference seal as boundary separating the two volumes from any contamination exchange. One or more conduits inside the capsule define passageways for the conveyed fluids. In a further embodiment the fluid conveyance device is a passive fluid conveyance device.

The described active diaphragm membrane pump relay on sensors and controls which allow the control unit to know the exact position of the membrane at any time. This feature allows complicated programming of the actual pumping performance. According to a further embodiment a non controlled, passive diaphragm membrane pump comprises a low cost solution when operating in tandem with an active diaphragm membrane pump. Reference is made to FIG. 3. The passive fluid conveyance device simplifies the column design and reduces the overall price. The passive membrane pump containing capsules gas pressure chamber is pre-pressurised (low pressures such as 0.01-1 bar) and will in passive stage force the elastic membrane inwards the gas chamber housing part.

In a further embodiment a further function is selected from a passive fluid collection container. The liquid expansion chamber is not pressurised and positioned with a liquid head (such 0.5-10 meter) relative to the column via conduits and tubing defining fluid passageways. Gravity becomes the driving force for liquid return into the column. Reference is made to FIG. 4.

When the bioreactor apparatus is in use and produces a biologic product a liquid nutrient media will be flowing between the capsules in the column with micro organisms present in the cultivation capsule(s). Typically, the top capsule and bottom capsule are in operable and liquid contact with each other and capable of regulating liquid to and from said bioreactor apparatus.

In a further embodiment of the purification apparatus at least two of the capsules are separated by a sealing arrangement creating a boundary between said two capsules creating an internal sterile environment further separating the two capsules internal functional devices against the external non sterile environment. In a preferred embodiment the sealing arrangement is a gasket. Such a gasket may be prepared from an elastic organic material, such as rubber or a polymeric material with hardness ranging from 20-90 shore A. Typically the gasket is an inflatable gasket containing a closed volume to receive a pressurised fluid through a connection for expansion of the flexible gasket. According to the invention the capsules corresponds with the neighbouring capsules in said column preferably via internal conduit tubes, conduit channels, fluid communication bus, power supply, or data communication, such as a power supply bus and a data communication bus axially from top to bottom via connections in each capsule. According to the invention said capsules comprises inter engage complementary mechanical connectors on each end-face for assembling purposes. Each capsule comprises one male and one female end-face side.

According to the invention the capsules corresponds with the neighbouring capsules in said column preferably via internal conduit tubes, conduit channels, fluid communication bus, power supply, or data communication, such as a power supply bus and a data communication bus axially from top to bottom via connections in each capsule.

According to the invention said capsules comprises inter engage complementary mechanical connectors on each end-face for assembling purposes. Each capsule comprises at least one mechanical connector on the capsule first connection surface and for the capsule second connection surface at least one arrangement selected from the group of:

-   -   one male connector     -   one female connector     -   no connector

Said capsules may be engaged and locked together by at least two methods:

-   1. At least one “floating” fast operation, quick engage/release,     snap locking circumference ring element. The radial extended portion     of the capsule circumference is a collar and bearing supporting the     partially rotation snap locking ring in axial direction thus able to     comprehend axial forces clamping two capsules towards each other.     The angled seat correspond with opposite orientation angled seat on     the neighbouring capsule and by rotating said snap locking ring     between 10 and 180 degree, preferably 30-45 degree, the two capsules     are forced towards each and locking mechanically together -   2. At least one “non floating” fast operation, quick engage/release,     clamp locking inner round and/or outer circumference oriented     device(s). The radial extended portion of the capsule inner round     and/or circumference is also a collar and a snap locking ring able     to comprehend axial forces clamping two capsules towards each other.     The (angled) locking grip (male part) correspond with opposite     orientation (angled) seat(s) on the (female part) neighbouring     capsule and by rotating said two capsules in opposite directions,     between 10 and 180 degree, preferably 30-45 degree, the integrated     snap locking devices insures the two capsules are engaged towards     each other locking mechanically said two capsules together

Any of the above assembling methods further generate fluid tight sealing either by:

-   1. compressing of at least one flexible, solid annular gasket(s) in     between two capsules for fluid tight connection, such rubber gaskets     preferably have shore A hardness ranging 5-95. Such as silicone     rubber with available range from 20-80, latex based rubber with     available range from 20-60, EPDM rubber with hardness from 40-90 -   2. expansion of at least one inflatable gasket between two capsules     serving two tasks; 1. by applying fluid pressure to an internal     volume the gasket increases thickness forcing the two capsules in     opposite directions until the snap locking element(s) are completely     engaged, 2. the inflatable elastic gasket seals the various     connections. The method of applying inflation pressure may be     applied to one capsule individually or all capsules in one operation     for a fast and secure assembling. Fluid connection from all capsules     inflatable gaskets may be assembled in longitudinal arranged     manifold channel within the capsule or in connection with the     circumference of said capsule

At least one such rigid or inflatable gasket preferably seals more than one fluid communication connections in order to avoid leakage or undesired fluid mixing or access to/from the environment. Said gaskets seals gas distributions channel and/or conduit port(s), primary liquid conveyance channel and/or conduit port(s), secondary liquid distribution channel and/or conduit port(s), fluid communication channel port(s), electrical distribution, communication ports or the like.

Said rigid or inflatable gasket may be used in combination between two capsules, such as one or more rigid gasket and one or more inflatable gasket with different sealing purpose and properties.

Furthermore the inflatable gasket expands by pressurization by a pressurised fluid being added to the internally volume. Said fluid may be a reaction curing fluid which alter the internally volume elasticity or increase the internally volume hardness.

In a further embodiment the further up-stream function is a conditioning capsule for controlling the temperature. A number of different designs of heat exchangers for cooling or heating, which are commercial available for temperature regulations, may be used and incorporated into the bioreactor apparatuses of the present invention in order to control the temperature of the liquid flowing through the parts of or the entire bio factory apparatus.

In a further embodiment the further up-stream function is a conditioning capsule for controlling pH and gas content, Bacterial cultures are aerated predominantly with air, cell cultures are aerated with multiple gases such as oxygen, nitrogen, CO₂, and air in a mixture, which is specific for each type of micro organism and the specific cultivation process. Gas composition is regulated by sensitive control systems, and an optimum gas atmosphere should be formed and maintained for the cultivation and manufacture of the product. During cell proliferation the pH of the medium changes and keeping pH constant is vital for many production processes and therefore it must be controlled. pH control are done by adding acids and/or bases in bacterial fermentation, but preferably cultivating mammalian cells with the addition of CO₂ controlling the pH. One or more capsule inlets are suitable for gas introduction.

In a further embodiment the further up-stream function is a fluid collecting container for fluid storage. An independent gas volume inside a capsule when covered with a membrane or a cylinder of elastic material allow liquid volume expansion or liquid pressure pulses to be accumulated eliminating damages to the column or the micro organisms.

In a further embodiment the further up-stream function is a manifold capsule. The manifold capsule facilitates support for both or either of:

-   -   one or more valves facilitating fluid control inside the column         periphery to one or more connected neighbouring capsules via         conduits     -   one or more valves facilitating fluid control via conduits to         external radial oriented ports, said external connections         comprise at least tubes for hose attachment or tri-clamp flanges

As an example, the two capsule covers both comprise tri-clamp flanges arranged identical on the inside surface of said capsules facing towards each other. Suitable oriented for being inter connected with commercial available size exclusion or chromatography cartridges, modules available from companies like Sartorius, Pall, Millipore, Meissner, etc. via the tri-clamp arrangement and process operating in parallel. Reference is made to FIG. 9. Said capsules may integrate one or more commercially available standard cartridges operating in parallel. One or more commercially available standard pleated cartridges operating in parallel. One or more commercially available standard plate&frame cartridges operating in parallel. One or more commercially available standard stacked disc cartridges arranged on top of each other and operating in parallel.

In a further embodiment the further up-stream function is a valve capsule. Relevant valves for integration with a capsule are pneumatic and/or electrical operated valves. Control of said valves being of proportional and/or on/off type of operation. Valves correspond via conduits in said capsule with selectable functions or devices within the invented column or external devices.

In a further embodiment the further up-stream function is a sensor integrated in one or more capsule(s). Critical process parameters that are often monitored include; pressure, pH, dissolved oxygen, conductivity, UV absorbance, flow, and turbidity. The packages that contain the traditional technologies for monitoring these parameters are not usually compatible with or effective when integrated into single-use assemblies for many reasons: cost, cross contamination, inability to maintain a closed system, and system incompatibility with gamma irradiation.

Typically, important sensor information is supplied from sensors such as; pressure sensors, temperature sensors, stain-gage sensor, flow sensors, glucose sensors, lactate sensors, pH sensors, oxygen sensors, CO₂ sensors, N₂ sensors, displacement sensors, proximity sensors, positions sensors, conductivity sensors, micro organism activity sensors, micro organism number counting sensors, cell mass sensors, etc. Preferably sensors are of single-use capability.

In a further embodiment the further up-stream function is an end cover capsule. A mechanical support capsule comprising complementary mechanical connectors on one end-face only for assembling purposes. The opposite side facilitates arrangements to transfer the column weight to a horizontal support. The columns can be anchored to the floor or placed on casters for easy mobility or placed on a bench-top. Its compact vertical design (up to six feet/two meters high) reduces floor space requirements. The system offers two manifold configurations bottom in/top out and bottom in/bottom out.

In a further embodiment the further up-stream function is an adaptor for different diameter capsules. Two different diameter or different cross section capsules may be inter-connected. Comprising two complementary mechanical quick engage/release couplings devices with different diameter or cross section integrated one on each side end-face of the same capsule. Said adaptor capsule redirects internally necessary connections between the possible different locations as to diameter or cross section.

In a further embodiment the further up-stream function is a power supply capsule. Preferably the bioreactor apparatus is an independently operating apparatus including all the benefits of the modern PLC (Programmable Logic Control) for control purposes. Such PLC excludes fully any needs of control from the commonly known PC (Personal Computer). The bioreactor apparatus further includes common supply of electrical power, of pressurised air, if desired of a vacuum source and if desired temperature conditioning fluids.

In a further embodiment the further up-stream function is a data acquisition capsule. The collection of sensor signals from at least one or more capsule is important for the process optimisation and performance.

The control device circuit board may hold at least one micro processor with an associated program and working memory for storing algorithms, programming, and various power supplies, regulating power output channels in order to control among others by means of communication:

-   -   Gas supply inlet valve(s)     -   Gas pressure outlet, exhaust valve(s)     -   Fluid control valve(s)     -   Proportional power control of various fluid conveyance devices     -   Proportional power control for temperature conditioning purposes

The regulating functions may be based on non-linear, adaptive, fussy logic, slide mode control, PI(D) regulation and combinations hereof. The sequence of events or regulation loops preferably in a hieratic set-up with internal loops avoiding external loops with individual data sampling speed being in the range of 0.1 to above 100 kHz. Compared to analogue operation, a digital signal bus is preferred as to its unlimited capabilities. Several industrial standards (such as Profibus, Fieldbus, Canbus, Modbus, etc) are able to control the bio factory apparatus through Ethernet, wireless network, RF or power-line-communication connection.

In one embodiment said capsule containing the electronics for data acquisition may integrate display(s) on the outer wall informing personal to easy input of information in parallel with external displays and monitors.

According to the invention the capsule to capsule correspondence is predominantly performed by internal conduit tubes, conduit channels, connectors, connections attached to or exposed to at least one capsule with the interface to external communication, such as an end-face capsule. The communication are ready to start when two opposite male/female end-faces inter engage with complementary mechanical connectors and the preferred aseptic film seals are removed and the desired gasket insure tightness the column is ready for operation.

Capsule to external devices fluid connection are facilitated by the radial oriented hose and/or tri-clamp connections extending from the capsules and it becomes possible to promote fluid connection externally. Such as mounting commercially available separation cartridges axial oriented as satellites around the column extending slightly from the column. Integration of commercial products facilitates reduced investment and broadens further supplier and products selection.

According to the invention the capsule body comprises the combination of one or more body parts manufacturing from identical of different organic materials creating the capsules for single-use applications.

-   -   Synthetic polymers in general, such as; polycarbonate,         polypropylene (PP), polyethylene (PE), PTFE, polyethersulfone,         polyethylene terephtalate/polyester

(PET), polyoxyethylene, polyacrylonitrile, Polysulfone, Polyethersulfone, Polystyrene, Polyacrylicsulfone, polystyrenes, ethylene vinyl acetate, cellulose acetate, polytetrafluoroethylene, Polychlorotrifluoroethylene, Polyfluoroethylenepropylene, polyvinylidene fluoride, polyvinylidene chloride (PVC), Polyamides, polycarbonate, methacrylic resins, acetal, acrylics, nylon, ABS and further copolymers thereof, resins in general and thermoplastics in general or blends thereof

-   -   Synthetic elastomers in general, such as silicone, EPDM, Viton,         neoprene, polyurethane, butyl     -   Natural polymers (also bio polymers) in general, such as rubber         and latex and any form of fibres based on cellulose such as:         paper, cotton, wool, flax, hemp, coconut, jute, viscose rayon     -   Synthetic bio degradable polyester (bio plastics) in general,         like; aliphatic polyester such as polylactic acid polymer         families and polyhydroxyalkanoates families, polybutylene         succinate families, etc     -   Carbon fibre and synthetic carbon fibre based substances         (Kevlar) and further carbon and carbon based structures in         general     -   Metal powders and/or metal fibres     -   Glass and/or glass fibres

or any combination hereof. The materials selected having translucent, clear or light impermeable properties. Polymers with low extractables and leachables, with the ability to accept high temperature sterilization and gamma sterilization being preferred. The possible partial use of non-organic materials for re-enforcement in organic mouldable materials is fibres such as metal, carbon and glass fibres. Manufacturing of at least one capsule body part predominantly performed by casting, moulding such as pressure casting, injection moulding where the selected, selections of raw materials under applied pressure and temperature flow into a mould which shape the body part. Larger bodies may benefit from being manufactured via rotation moulding techniques.

Capsule parts material selected as to the operating pressure and the preferred polymers properties which my further comprise strengthening by the addition of, inclusion of organic fibres like carbon, Kevlar, plant and cellulose or inclusion of in-organic fibres like glass or metal fibres into a composite type of material.

For permanent use the capsules may be traditionally manufactured from solid metals. Capsules for single-use and permanent use may be combined in the same column.

According to the invention aseptic assembling of two or more capsules into a column is promoted by—peel strip or barrier film covering openings, connection openings, pipes, tubes, channels etc with access to the capsule internals.

Breakable aseptic seal(s) in film or foil may cover at least parts of the capsules end-face surface. Breakable seals in foil are covering each individual through going, fluid connector, manifold and flange for interconnections and protects the surface of and internals of the capsules from the surroundings. The capsules are stacked and the integrity seals are removed by pulling the seal in a radial direction exposing the two fluid connections to each other ready for interfacing. Use of seals on capsules is relevant on both bench-top systems and larger floor mounted systems. Bench-top systems could alternatively be delivered pre-assembled from the supplier.

Two capsules end-face foil seals facing each other protect a sterilized and gas protected surface, internal devices and connections. The foil is double in a design where the outer foil is equipped with a handle and a 180 degree bend opposite of the handle. When the handle is pulled the sealing foil will slip the circumference attachment. When two capsule seals are removed simultaneously no contamination reaches the fluid connection and gas connection. Also a slight overpressure of protective gas in each capsule insures no contamination is sucked into the capsules.

A thin foil is integrated and glued on the circumference for protection purposes, and cover the entire end-face and all the fluid flanges. Each capsule is to be connected prior to rotation to the next capsule controlled by pairs of male and female locking devices located in symmetrical mode on the capsule end-face outside the cover seal, which allow capsules of the column to be assembled without breaking the seals and further prohibit incorrect orientation. The foil is to be broken when two capsules are correctly positioned towards each other exposing fluid connection, conduits, gas connection, electrical connectors rotational correct oriented towards the neighbouring capsule. The foils are removed simultaneously by dragging both foils of in a double mode configuration without exposing any end-face to the local environment. After seal removal the two capsules are locked together for positive connection via the inter engage complementary mechanical connectors on each capsule end-face finalising the assembling.

Yet an aspect of the present invention utilising the option of the functional devices supplied pre-enclosed and pre-sterilized in a plastic film bag prior to integration into said capsule concept. Functional devices, such as cultivation functions, size exclusion functions, polishing functions, separation functions, ion exchange functions and chromatography functions may benefit from such assembling methods.

The functional device supplied in a film bag combines the film properties, being a thin foil, with the advantage of a breakable seal arrangement in said bag film wall. Said bag utilise the incorporation of fluid conveying and breakable sealing arrangement for fluid conduit, connection purposes to the neighbouring functional device(s) vastly eliminating contamination problems. During assembling into a column the breakable seals are opened allowing fluids to pass.

The described method of assembling functional devices supplied pre-packed in film bags and pre-sterilized allow the use of capsules for more permanent use. Such as capsules of re-usable character which by sterilisation obtain yet a life of use. Such capsules may be manufactured from metals hereby obtaining properties not easily obtainable from polymer based capsules such as high pressure usage.

In a further aspect the present invention concerns a purification apparatus having a top capsule and a bottom capsule, the apparatus comprising a processing capsule having a purifying device for purifying a biologic product, and a second downstream capsule, wherein said capsules are stacked and locked in a column, with the proviso that the top capsule or bottom capsule are not pump capsules.

Thus, the top capsule may be a processing capsule having a purifying device and the bottom capsule may be the second downstream capsule or may be the third or so forth capsule. Alternatively, the processing capsule may be located between the top and bottom capsules and may consist of one or more processing capsules.

The purification apparatus is preferably for single use, or is pre-sterilized or both. The purification apparatus may be any suitable apparatus, such as a down-stream function selected from separation, filtration, chromatography, or ion exchange.

The purification apparatus may be connected to a bioreactor apparatus according to the present invention or may be connected to any prior art bioreactor apparatus such as those disclosed in the background section. After manufacturing a biological product, the crude product is transferred to the purification apparatus of the present invention to be processed into a pure biological product.

In a further embodiment the purifying device is in the top capsule.

In a further embodiment an external fluid conveyance device, such as a peristaltic pump, is connected to the column.

In a further embodiment the purification apparatus comprises liquid media and the biologic product to be purified.

In a further embodiment the second down-stream capsule is selected from a separation capsule, filtration capsule, chromatography capsule, ion exchange capsule, conditioning capsule, sparging capsule, diffusion capsule, active fluid conveyance capsule, passive fluid conveyance capsule, fluid collecting container, valve capsule, column end capsule, manifold capsule, storage capsule, power supply, data acquisition and control capsule. In a further embodiment the, purification apparatus comprises a further down-stream function selected from separation, filtration, chromatography, ion exchange, conditioning, sparging, diffusion, active fluid conveyance, passive fluid conveyance, valve, column end, manifold, storage functions, heating, cooling, power supply, data acquisition and controlling wherein the down-stream function or functions are located in one or more capsules.

It is to be understood that some of the down-stream functions, such as functions in one or more capsules, such as conditioning, sparging, diffusion, active fluid conveyance, passive fluid conveyance, valve, column end, manifold, storage functions, heating, cooling, power supply, data acquisition and controlling, have the same meaning as defined and explained above in relation to description of the bioreactor apparatus up-stream functions, and it is intended that such functions also apply as embodiments of the purifying apparatus of the present invention.

The Down-Stream processes are typically:

1. Clarification, purification with membrane filters 2. Sterile filtration with membrane filters 3. Polishing with membranes 4. Polishing with resins/sorbent based chromatography

Polishing based on chromatography is a complex task involving various methods and products. Such as the traditional packed bed consisting of a resin operating as in single product or mixed product mode. Or the modern membrane based chromatography, such membranes mixed with resins and/or charged by product or external means. Flow path being continuously or sequentially with a holding, retention time. Or all the methods and products are mixed. It is not the scope of the present invention to enter the field of chromatography, but to include and integrate any of the disposable chromatography products and methods into the invented disposable bioreactor and/or purification apparatus.

Some embodiments of the invention provides a single-use bioreactor and/or purification apparatus, also known in the industry as process train, each pre-sterilised capsule corresponds in an aseptic way with at least one neighbouring pre-sterilised capsule and is loaded directly from the previous capsule, thus circumventing multiple and lengthy connection hoses, clamps, connectors, external valves, peristaltic pumps connected with bio container, tanks eliminating cost and possible cross contamination. Further better utilizing floor space, decreasing process time, and allowing for more flexible Down-Stream purification flow path. As one result, there's no need for CIP/SIP, a step that can consume as much as one third of the total process time.

In a further embodiment of the purification apparatus at least two of the capsules are separated by a sealing arrangement creating a boundary between said two capsules creating an internal sterile environment further separating the two capsules internal functional devices against the external non sterile environment. In a preferred embodiment the sealing arrangement is a gasket. Such a gasket may be prepared from an elastic organic material, such as rubber or a polymeric material with hardness ranging from 20-90 shore A. Typically the gasket is an inflatable gasket containing a closed volume to receive a pressurised fluid through a connection for expansion of the flexible gasket. According to the invention the capsules corresponds with the neighbouring capsules in said column preferably via internal conduit tubes, conduit channels, fluid communication bus, power supply, or data communication, such as a power supply bus and a data communication bus axially from top to bottom via connections in each capsule. According to the invention said capsules comprises inter engage complementary mechanical connectors on each end-face for assembling purposes. Each capsule comprises one male and one female end-face side.

In a further embodiment the purification apparatus comprises a size exclusion capsule. According to the present invention, the capsule concept integrates a variety of porous materials defined as membranes or depth filter based on a variety of materials and configurations. A capsule may integrate one or more commercially available standard cartridges operating in parallel. One or more commercially available standard pleated cartridges operating in parallel. One or more commercially available standard plate&frame cartridges preferably operating in parallel.

One or more commercially available standard stacked disc cartridges arranged on top of each other and operating in parallel. In such embodiment two individual capsules end-faces both comprise connection arrangements on the inside surface of said capsules facing towards each other with a distance suitable for being inter connected with commercial available cassettes and/or cartridges available from companies like Sartorius, Pall, Millipore and operating in parallel. Orientation may be axial as well as radial or combinations hereof. In some applications the embodiment need no cylindrical device interfacing the two capsule end-faces. In other application its advantageous to enclose the integrated separation devices inside a cylinder fluid tight sealed with the two capsule end-faces.

According to an embodiment of the invention, the closed capsule integrates a pleated filter material. A pleat is a type of fold formed by doubling fabric, sheets backed upon itself and securing it in place. Pleating is a compact method of obtaining high loads of fabric, porous sheets and used extensively in the filtration industry and the manufacturing method is often called knife pleat. If order to avoid each pleat gets in contact to the neighbour pleat separators, spacers with fluid capability are arranged between the pleats. The pleat membrane thickness is selected in order to get the desired flow and pressure drop over the membrane. The pleat will typically be of the same thickness all through the core. The pleat design may be with straight pleats or curved pleats for higher surface area. Two or more individual diameter pleated bodies may be arranged into the same housing with one pleated body surrounding the other with the upstream volume shared, being in between the two bodies so one downstream becomes the centre of the in diameter smaller body and the other downstream the circumference of the in diameter larger body.

According to an embodiment of the invention, the closed capsule integrates at least one spiral wound filter element of cylindrical and elongated design with spiral oriented layers of non-woven sheet. Said sheet laminating drainage channel spacers on one side and on the other side the feed channel spacers all wound around a central perforated fluid collection tube. The drainage spacer is shorter and less wide than the non-woven sheet and on three sides sealed from the feed stream with seals between and/or bonding of the non-woven sheet edges, which hereby form an envelope around the drainage spacer, though the drainage spacer is open to, connected to and in unhindered permeate fluid correspondence with the perforated fluid collection tube. The feed spacers are open on all four edge sides and with no fluid connection to the fluid collection tube receiving the permeate. Anti-telescopic members are mounted at both end surfaces of the sandwich circular core.

According to a further embodiment of the invention, the capsule comprises two or more rotation symmetrical filter disc elements stacked inside the capsule. Said elements comprise two porous discs with central inlet and closed at the circumference creating the outlet from the external surface of the element. Such filter discs are commercially available from companies such as Begerow, Cuno and Filtrox.

Media is typically mixed in bulk and then aseptically transferred to the bioreactor. Pre-filtration is used to remove the bulk of particulate and colloidal contaminants from the media in order to extend the service life of the filter train. Pre-filters must be sized appropriately to handle batch to batch media variability, as well as ensuring that the sterile media fill in the bioreactor is completed successfully and on time.

In a further embodiment the purification apparatus comprises an ion exchange capsule. According to an embodiment of the present invention, an ion exchange (cation and anion) based membrane absorbers are relatively thin, synthetic, micro-porous, macro-porous membranes that are derivatized with functional groups similar to those on the equivalent resins. Each membrane absorbers are 5-30 layers of selected sheet material with the desired properties stacked laminated with flow guides in the capsule generating a much smaller foot print compared to bed columns. Membranes feature a more open structure than resins almost eliminating the diffusion limitations. The reduced size reduces the buffer consumption but increases the flow rate. Despites this phenomena adsorption is efficient because the transport of solutes to their binding sites in a membrane absorber occurs mainly by convection, while pore diffusion is minimal. These benefits reduce process time to less than 10% of those associated with traditional chromatography bead based columns. Said membrane absorbers have less than 20 mm thickness compared to traditional column bed high of more than 200 mm.

According to a further embodiment of the invention a set of two identical and mirrored capsules cover parts, a first capsule cover part and a second capsule cover part arranged opposed comprising one or more ports for axial internal integration of size exclusion or chromatography cartridge or ion-exchange cartridges. Two principles of said cartridges are available; 1. independent cartridge element encapsulated in its own housing, 2. independent cartridge element not encapsulated in its own housing and hereby depending on individual housing for each cartridge or sharing arranged in parallel mutually a housing. Reference is made to FIG. 9. Said standard cartridges supplied with standard connection ports. The invented capsule cover parts equipped with ports corresponding desired cartridges and selected from the group of:

-   -   a tri-clamps ports gasket sealed     -   hose connection ports     -   hollow tube ports for internal connection O-ring sealed     -   hollow tube ports for external connection O-ring sealed

Said set of two identical and mirrored capsule cover parts, a first capsule cover part and a second capsule cover part arranged opposed facilitates process capacity adjustment by blocking off sets of ports, one port on the first capsule cover parts and axial opposed a port on the second capsule covers part.

In further one variation said embodiment is arranged integrated within the column as the top functional capsule. Said variation with only one capsule cover part and the connected cartridges oriented axial in one cartridge connection end only facilitating both inlet and outlet in the same cartridge connection end hereby reducing the overall cost.

In a further embodiment the purification apparatus comprises a chromatography active resins or sorbent capsule.

According to a further embodiment of the present invention, polishing or product recovery in industrial scale is typically performed via chromatography by resins appearing as porous particles, beads or granular substance packed, contained in a bed structure of stainless steel and cylindrical translucent walls. The present invention integrates the use of chromatography active resins/sorbent shaped as porous, fibrous, beads particles or granular substance contained within a capsule. Said capsule may be fully single-use with its content or the content or capsule may be re-usable.

Polishing column or bed chromatography based on reusable particles or granulate washed after each use is traditionally performed in combined stainless steel and glass vessels with heavy gear motors connected via threaded rods to pistons for vessel volume changes. A setup far from disposable! The present invention allows the volume change performed inside a capsule by simple means. Such as at least one flat free floating piston or membrane with axial operation controlled by controlling fluid pressure on the side opposite to the chromatography media. Increased controlling fluid pressure reduces the media bed volume by piston movement. And opposite the decreased controlling fluid pressure increase the bed volume by piston movement. The piston movement velocity and variable displacement determined by the controlling fluid. The piston includes a circumference seal as boundary separating the two volumes from any contamination exchange. Bed height ranging from 100 mm to 1,000 mm and the diameters from 100 mm to 1,000 mm.

Improved packing by the resin being added in one or more steps to the capsule allowed to settle under the effects of gravity and then subjected to one or more treatments of vibration from the vibration devices until a suitably packed capsule is obtained. Liquid used to suspend the resin while being placed into the capsule may be at least partially removed before or during the vibration step(s).

Air contraption removed by connected vacuum or low pressure for venting. If needed sequential substance washing for regeneration and/or product recovery performed by fluids supplied by nozzles integrated in the capsule. Sequential or continues fluidising of said substances within a capsule improves micro organism or product removal and affinity based purification. Fluidising further improves the packing density of said substance. Capsule may be partly manufactured from translucent materials for visual inspection and improved platform programming.

In a further aspect the present invention concerns a bio factory apparatus assembly for producing a purified biologic product comprising a bioreactor apparatus and a purification apparatus in one column having a top capsule and a bottom capsule, the apparatus comprising a cultivation capsule having a production device for producing a biologic product, and a processing capsule having a purifying device for purifying a biologic product, wherein said capsules are stacked and locked in the column, with the proviso that the top capsule and bottom capsule are not pump capsules at the same time.

Such a bio factory apparatus assembly for producing a purified biologic product may typically be composed of the bioreactor of the present invention as described and explained above and the purification apparatus of the present invention as described and explained above.

It is understood that all aspects and embodiments in connection with the bioreactor apparatus and the purification apparatus described above are also aspects and embodiments in the bio factory apparatus assembly.

It should be understood that the present invention covers different aspects and embodiments, such as capsules of various functionality selected according to the product to be manufactured and stacked into a column. The bio factory apparatus operates in its life time on continues basis serving various tasks. Said platform comprises individual capsules integrating at least one specific function and/or device, such as:

1. Cultivation of micro organism in bioreactors, fermenters 2. Conditioning of temperature, heating and/or cooling 3. Conditioning of pH and gas in general such as O₂, CO₂, N₂ 4. Separation by size exclusion 5. Purification by membrane chromatography and/or ion-exchange 6. Purification by resin/sorbent chromatography 7. Active fluid conveyance or active mixing, agitation 8. Passive fluid conveyance 9. Passive fluid collection container being also an independent embodiment 10. Fluid storage 11. Manifold for internal as well as external communication 12. Valve(s) for fluid control 13. Sensor(s) for condition measurements 14. End cover(s) for bottom support and/or top cover 15. Adaptor for different diameter or cross section capsules 16. Power supply, such as electricity, pneumatics, vacuum 17. Data acquisition, sensor interface, PLC control, communication

According to the invention the invented capsule concept comprises further the combination of at least two of any of the above listed different functions, devices or processing purposes into one single capsule. Combinations in one capsule comprises also such as:

-   -   separation via surface filtration and depth filtration     -   purification and surface/membrane separation     -   active fluid conveyance and valve(s)     -   Active fluid conveyance and bioreactor(s)     -   Active fluid conveyance and controlled bed volume chamber     -   Valve(s) and passive fluid expansion     -   End cover and fluid expansion or fluid collection     -   End cover and manifold     -   Separation/purification and fluid conveyance     -   Controlled bed volume and fluid conveyance     -   End cover and power supply and/or data acquisition     -   Manifold and valve(s)

The combinations are limitless though not limited in any way or number to the above.

In a further aspect, the present invention concerns a method of preparing a biologic product comprising the steps of:

-   -   Cultivating a micro organism in a bioreactor apparatus of the         present invention, and collecting the biological product. The         bioreactor apparatus may be selected from or composed of any one         of the above embodiments as well as from any one of the specific         embodiments shown in FIGS. 1-10. The biologic product may be         selected from any one of the products as defined herein, which         constitutes embodiments of the invention. The collection of the         biologic product may be done according to prior art means or may         be done in accordance with the purifying apparatus of the         present invention.

In a still further aspect, the present invention concerns a method of purifying a biologic product comprising the steps of:

-   -   Applying the un-purified biological product to a purifying         apparatus of the present invention, and collecting the         biological product in purified form. The purifying apparatus may         be selected from or composed of any one of the above embodiments         as well as from any one of the specific embodiments shown in         FIGS. 1-10. The biologic product to be purified may be selected         from any one of the products as defined herein, which         constitutes embodiments of the invention. The previous         cultivation and production of the biologic product may be done         according to prior art means or may be done in accordance with         the bioreactor apparatus of the present invention.

In a further aspect, the present invention concerns a method of producing a purified biologic product comprising the steps of:

-   -   Cultivating a micro organism in a bio factory apparatus assembly         of the present invention, and collecting the biological product         in purified form. By combining bioreactor apparatus of the         present invention with the purifying apparatus of the present         invention a complete bio factory is assembled in one column.         Obviously, all embodiments of any one of the bioreactor         apparatus and purification apparatus applies to this invention         as well.

DETAILED DESCRIPTION OF THE FIGURES

FIG. 1.

Bio factory apparatus (as claimed in claim 1) stacked and locked in a column 10 with external mounted independent active fluid conveyance pump 19. Said stand-alone active fluid conveyance pump 19 (containing one or more valves for desired functionality) is oriented next to the column 10 and fluid vice via hoses 19 a, 19 b connected to one or more the tri-clamp ports 11 on said column 10. The column 10 is composed of a selection of capsules comprising a first end-face/manifold/valve capsule 12 a supporting the vertical column 10 on a horizontal surface. A second size exclusion capsule(s) 13, a third manifold/valve capsule 14, a fourth cultivation capsule 15 hosting the micro organism, a fifth end-face/manifold/valve capsule 12 b topping of the column 10 containing one or more valves 17 b. The external pump 19 is connected to bio containers 18 a and 18 b, distributing fresh media to the column 10 and delivers processed product collected in second bio container 18 b after purification. Said pump 19 comprises a substantial variation in mass capacity servicing the requirements from the individual functionality capsules. Further the ability to service different tasks as to the selected sequence for opening and closing the valves behind tri-clamps 11 in or on valve 17 said column. Centrally mounted double valve body 17 controls the mass flow in both directions between manifold/valve capsule 14.

Said purification capsule 13 comprises sets of stacked porous discs 13 a, 13 b separated by first inlet fluid flow guides/spacers 13 c and second outlet fluid flow guides/spacers 13 d, said first inlet fluid guide/spacers 13 c in fluid contact only with supply, central volume 13 e and said second fluid guide/spacers in contact only with drainage, circumference volume 13 f.

Said cultivation capsule 15 comprises sets of stacked porous discs 15 a, 15 b separated by first inlet fluid flow guides/spacers 15 c and second outlet fluid flow guides/spacers 15 d, said first inlet fluid guide/spacers 15 c in fluid contact only with supply, central volume 15 e and said second fluid guide/spacers in contact only with drainage, circumference volume 15 f.

Forced flow direction for purification capsule 13 and cultivation capsule 15 as desired either as distributed from the central volume radial along the fluid guides—passing the discs perpendicular—along the fluid guides radial for collection at the circumference volume or vice versa.

Tri-clamp flanges 11 not in use are blocked off.

FIG. 2.

Bio purification apparatus (as claimed in claim 13) stacked and locked in a column 20 comprises a selection of capsules for purification purposes comprising a first end-face/control capsule 21 supporting the complete vertical oriented column 20 on a horizontal surface. A second manifold/valve capsule 22, a third size exclusion processing capsules 23, a forth pump control capsule 24, a fifth active fluid conveyance pump capsule 25, a sixth size exclusion processing capsule 26, a seventh polishing processing capsule 27, a eight end-face/manifold/valve capsule 22 b topping of the column 20. Said internal active fluid conveyance pump 25 a in pump capsule 25 conveys and exchanges fluid along said column 20 to and from one or more capsules via internal and/or external conduits and further with the associated bio containers 28 a, 28 b via valves in manifold/valve function capsules 22 a, 22 b, 26 and pump capsule 25. Two centrally mounted double valve bodies 28 controls the mass flow in both directions axial in the column 20. The first end-face/control capsule 21 controls all the valve functions, the pump function and collect data for distribution via the communication and power supply bus and further communication with operator. Said capsule 23 or 27 may be equipped with a display for on site visible information.

The column may be oriented horizontal on a surface or in a supporting rig if desired.

FIG. 3.

Bio factory apparatus (as claimed in claim 19, 20) stacked and locked in a column 30 comprises a first capsule 31 integrating the overall column control, internal as well as external communication with the column, integrated into the end-face capsule 31. An active fluid conveyance device 32 a operates as the active membrane pump in a second capsule 32 integrating valve functions (not visible). A third capsule 33 integrates the purification function. A forth capsule is a combined manifold/valve capsule 34 a integrating one or more valves 34 b and further conduits corresponding with tri-clamp flanges 34 c connected via hoses (not shown) to at least external containers (not shown). A fifth cultivation capsule 35 integrates the micro organism cultivation function. A sixth capsule 36 being a manifold/valve capsule, seventh capsule 37 integrates a passive fluid conveyance device, such as a passive membrane device 37 a operating as a collection and expansion container further integrated within the top capsule 37 of the column. The diaphragm pump capsule 32 has a membrane 32 a which oscillates controlled by the variation in drive gas pressure on the dry side 32 b. Said membrane 32 a corresponds with the fluid to be conveyed on the opposite and wet side 32 c. The seventh capsule 37 has a closed chamber 37 b which is pre-pressurised and will in passive stage force the elastic membrane 37 a towards the chamber 37 b in capsule 37. In one of many principle methods of column operation the pumped fluid conveyed at low pressure from second capsule 32 pass the fifth cultivation capsule 35 to the seventh capsule 37 fluid chamber 36 c. Hereby the gas chamber 37 b becomes pressurized according to the liquid volume the active pump 32 is conveying. When second active capsule pump 32 having membrane 32 a is fully expanded a corresponding liquid volume is stored in the passive pumps wet chamber 37 c. As soon as the second pump capsule 32 membrane 32 a deflects as to removed drive gas pressure the stored liquid will return passing the desired capsule(s) in column 30.

One centrally single valve body 38 controls the fluid access to and from the passive pump capsule 37 fluid, wet side 37 c.

One or more centrally located dual valve bodies such as valve body 39 allow and control the fluid direction being conveyed axially in the column. Inner valve body 39 a corresponds with the central volume of the column and the outer valve body 39 b corresponds with circumference volume of capsule 33. Said valve body comprises two individual operated circular elastic valve bodies 39 a, 39 b corresponding with conduits 77 a and 77 b in capsule covers allowing fluids passing simultaneously in either direction or in two different directions without being mixed. Said two individual elastic valve bodies comprise an inner valve element 39 a and circumference valve element 39 b. Valve 39 c controls the length of central volume 39 d extending from capsule 32 pump to capsule 37 collection chamber. Numerous valves similar to 34 b allows for hose connections external of the column for desired purposes and fluid conveyance. In one of several principle methods of column operation the pump membrane 32 a convey a desired volume from fifth cultivation capsule 35 passing forth capsule 34 valve function through the third capsule 33 without contact with the purification element 33 a at low pressure. Appropriate valve(s) in forth capsule 34 is closed/opened and the desired volume collected in wet chamber 32 c of pump 32 are free to set under pressure the desired volume passing the third capsule 33 purification unit 33 a.

FIG. 4.

Bio factory apparatus (as claimed in claim 19, 20) stacked and locked in a column 40 comprises a first pump control capsule 41, second capsule 42 integrating an active fluid conveyance device, such as a diaphragm pump membrane 42 a, a third capsule 43 integrates a filter for the size exclusion process, a forth capsule 44 combines manifold and two different valves 44 a, (reference is made item 39 in FIG. 3) 44 b, a fifth capsule 45 integrates a cultivation device 45 a, a sixth capsule 46 integrates end cover, manifold, one valve type with tube connection 46 a, 46 b to above the column 40 mounted external fluid collection container 47. Said fluid collection container 47 with access to the atmosphere through a sterile filter (not shown). The diaphragm pump membrane 42 a operates oscillating with rear side membrane 42 b pressure variation from atmospheric pressure to maximum 5 bar pressure. The membrane oscillates controlled by the variation in drive gas pressure on the dry side and the driven fluid on the opposite wet side. The liquid collection chamber 47 is not pressurised and positioned with a liquid head (such as 0.5-20 meter) above the top of the column 40. The pumped fluid conveyed from second capsule 42, containing the diaphragm 42 a passes optional, in one sequence the fifth capsule 45 containing the production device 45 a (according to actual column design) said fluid lifted further into the fluid collection container 47 against the force of gravity. Hereby the fluid collection container 47 becomes filled according to the liquid volume the active pump is conveying and relative to the liquid head. When the second capsule 42 containing pump membrane 42 a is fully expanded a corresponding liquid volume is stored in fluid collection containers 47. As soon as the second capsule 42 a membrane becomes drive gas pressure less the stored liquid will return passing the desired capsule(s) or other device. Other pumping principles may be chosen substituting the example membrane pump. Only one bio container 48 is shown for simplicity.

FIG. 5.

Showing a part of the bio factory apparatus. The fluid conveyance first capsule 51 comprises a centrifugal pump wheel 52 a driven by an electrical pancake motor 53 a integrated in a separate second capsule 53. The centrifugal wheel 52 a with vanes 52 b rotates in a round flat circular chamber 52 c with at least one central low pressure fluid inlet 52 d and high pressure fluid collection chamber 52 c at the circumference of said pump wheel. Said pump wheel 52 a fitted to a vertical axle 52 e corresponding through a rotating seal 52 f and a bearing 52 g in the upper pump chamber 52 c wall 53 b via the coupling 53 d associated with electrical motor 53 a mounted in its own capsule 53.

FIG. 6.

Showing a part of the bio factory apparatus. The fluid conveyance device in a first capsule 61 comprising one or more radial oriented valves an axial centre valve, a second capsule 62 comprising one or more radial valves and the centrifugal pump driven by an electrical device 63 a housed in a third capsule 63. The centrifugal pump wheel 62 a with vanes 62 b rotates in a round fluid filled flat chamber 62 c with at least one central low pressure fluid inlet 62 d and at least one circumference high pressure fluid outlet volume shared with chamber 62 c further integrated in said second capsule 62. Said pump wheel 62 a fitted to a vertical axle rotating aligned by one or more fluid lubricated bearing 62 e. Said pump wheel 62 a integrates one or more permanent magnets 65 a, 65 b, 65 c (creating the rotor of the drive unit) said magnets 65 a, 65 b, 65 c facing pump chamber wall 63 b in the third capsule 63. Said third capsule 63 comprises the electrical device 63 a part of the electrical drive unit inside capsule 63 which comprises at set of metal wire coils 66 a, 66 b, 66 c radial oriented symmetrical around the pump wheel 62 a rotating centre. The first disposable capsule 61 and the second disposable capsule 62 and the third reusable capsule 63 attached mechanically to each other, by the inter engage complementary mechanical connectors, creating the fluid conveyance device. Fluid connections may be performed in radial through valves/flanges 67 or axial direction through valve 68 in any number and arrangement.

FIG. 7.

Showing parts of the bio factory apparatus. The invented column comprises capsules interfacing with two mirrored capsules covers 71 a, 71 b and connected with the circumference cylinder 71 c in between said capsule covers 71 a, 71 b creating a volume 72 occupied by one or more of the functional devices. The capsule design inter connection is performed via the capsule integrated inter engage complementary mechanical connectors 75 a, 75 b comprising a set of quick engage/release clamps for locking the round inner and the round outer locking devices together. The (angled) locking grip (male part) 75 a correspond with opposite orientation (angled) seat(s) on the (female part) 75 b neighbouring capsule and by rotating said two capsule covers 71 a, 71 b in opposite directions, between 1 and 180 degrees. The integrated snap locking devices 75 a, 75 b locks mechanically said two capsules 71 a, 71 b together. Sets of conduits 77 a, 77 b in capsule covers 71 a, 71 b allow dual axial fluid transfer with the column and between capsules. Gasket 76 positioned between the two capsules covers 71 a, 71 b generate the sealing function insuring the two capsule covers 71 a, 71 b are constantly engaged, forced from each other and simultaneously sealing conduits, communication ports and connection. The sealing method comprises at least one annular inflatable gasket 76 oriented between two capsules covers 71 a, 71 b and by applying fluid pressure to an internal volume in said gasket the gasket 76 increases the thickness filling the volume between two sealing areas on two closed coupled capsules 71 a, 71 b. At least one such gasket 76 preferably seals one or more fluid connections, between said capsule covers 71 a, 71 b in order to avoid leakage or undesired fluid mixing, contamination or access to/from the environment. Connection inlet tubing 77 integrated in said gaskets 76 circumference extending from said gasket 76 radial to capsule 71 a, 71 b circumferences being accessible also after capsule interfacing.

FIG. 8.

Showing a part of the bio factory apparatus. In one embodiment the inflatable flexible annular gasket 86 sealing method comprises two gasket surfaces 86 a, 86 b facing in each axial direction. Inlet tubing 88 comprises a one-way valve 89 and a mechanical device for connection to an external device for inflation with a fluid under pressure. Gasket thickness increase from 0.5-25 mm non-inflated to increased thickness when expanded, such as 1-50 mm, though depending on the designed distance between two capsule cover sealing areas. The gasket facilitates fluids being conveyed through conduits 87 a, 87 b matching conduits or passageways in said capsule covers.

FIG. 9.

Showing a part of the bio factory apparatus. Set of two identical mirrored capsule cover parts, a first capsule cover part 91 a and a second capsule cover part 91 b, which both comprises one or more sets of tri-clamps ports 94 for axial integration of one or more commercial available purification cartridges 98 within the periphery of the capsule overall dimension. On the inner side of both capsule covers sets of tri-clamp flanges 94 (or alternative couplings) are arranged with integrated fluid correspondence via flat and circular chamber 95 to the capsule cover external fluid conduits or passageways 97 b passing gasket 96. Conduit 97 a is closed on this illustration.

FIG. 10.

Showing a part of the bio factory apparatus. Set of two non identical mirrored capsule cover parts, a first outlet capsule cover part 101 a and a second inlet capsule cover part 101 b. Both capsule covers comprises sets of dual purpose cylindrical push-in first female function tube ports 102 for internal and axial integration of commercial available purification cartridges with O-ring 102 a sealing. Said tube port 102 comprise external integration of tube pieces with O-rings sealing on the outside of said tube port 102 now with a second male function, said tube piece surrounds said cartridges securing the fluid inlet volume.

First outlet capsule cover part 101 a comprises sets of push-in female ports 102 arranged with common fluid correspondence via circular chamber 103 to the capsule cover external fluid conduits or passageways 107 b passing gasket 106. Passageways 107 a is closed, but used as fluid inlet conduit at the opposite second inlet capsule cover part 101 b. Second inlet capsule cover part 101 b comprises sets of push-in male ports 104 each with several fluid holes 104 a further with fluid correspondence commonly to circular chamber inside the capsule cover with access to external fluid conduits or passageways 107 a (not visible).

The standard elongated cartridges available from the industry are designed to fit only one cartridge into one slim line filter housing acting as the tube piece 109 surrounding the cartridge. Alternatively the individual tube pieces 109 may be replaced by one large diameter tube (not shown) extending from the first outlet capsule cover part 101 a to the second inlet capsule cover part 101 b. Diameter of said large tube equivalent to the capsule cover diameter.

Fluid flow pattern is; 1. Inlet in centre of casket 106 in second capsule cover, 2. Distributed in chamber inside capsule; 3. Distributed through heach of male tubes 104; 4. Through holes 104 a to volume outside the cartridges 108 and inside tube pieces 109; 5 forced through the cartridges 108 to the internal collection volume of said cartridge; further to the collection volume 103 and out through the capsule cover conduit circumference passageway 107 b.

The presented capsule cover part principle design as illustrated in FIG. 9 and FIG. 10 allow simple assembling of various commercial available size and principles of size exclusion cartridges or chromatography cartridge or ion exchange cartridges.

The FIGS. 1-6 do not include the invented gasket principle or other gasket principles. Such gasket principle has been left out for simplicity of the illustration.

It should be understood that each of FIGS. 1-10 illustrates embodiments of the invention.

This invention includes all modifications and equivalents of the subject matter re-cited in the aspects or claims presented herein to the maximum extent permitted by applicable law.

The present invention is further illustrated by the figures which, however, are not to be construed as limiting the scope of protection. The features disclosed in the foregoing description and in the figures may, both separately and in any combination thereof, be material for realising the invention in diverse forms thereof.

All methods described herein can be performed in any suitable order unless other-wise indicated herein or otherwise clearly contradicted by context.

The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise indicated. No language in the specification should be construed as indicating any element is essential to the practice of the invention unless as much is explicitly stated.

The citation and incorporation of patent documents herein is done for convenience only and does not reflect any view of the validity, patentability and/or enforceability of such patent documents. 

1. A bioreactor apparatus having a top capsule and a bottom capsule, the apparatus comprising a cultivation capsule having a production device for producing a biologic product, and a second upstream capsule, wherein said capsules are stacked and locked in a column, with the proviso that the top capsule and bottom capsule are not pump capsules at the same time.
 2. The apparatus of claim 1 wherein the production device is in the top capsule or bottom capsule.
 3. The apparatus of claim 1 further comprising an external fluid conveyance device, such as a pump means, connected to the column.
 4. The apparatus of claim 1 comprising liquid nutrient media and micro organisms.
 5. The apparatus of claim 1 wherein the top capsule and bottom capsule are in operable and liquid contact with each other and capable of regulating liquid to and from said bioreactor apparatus.
 6. The apparatus of claim 1 wherein the production device comprises a porous matrix or a matrix body for retaining micro organisms.
 7. The apparatus of claim 1 wherein the second up stream capsule is selected from a manifold capsule, cultivation capsule, active fluid conveyance capsule, passive fluid conveyance capsule, fluid collecting container, valve capsule, column end capsule, conditioning capsule, size adaptor capsule, power supply capsules, data acquisition capsules, control capsule and storage capsule.
 8. The apparatus of claim 1, comprising a further up-stream function selected from cultivation, separation, filtration, conditioning, sparging, diffusion, active fluid conveyance, passive fluid conveyance, collection container, valve, column end, manifold, storage function, heating, cooling, power supply, data acquisition and controlling, wherein the up-stream function or functions are located in one or more capsules.
 9. The apparatus of claim 1 wherein the active fluid conveyance capsule is an active reciprocating fluid conveyance capsule.
 10. The apparatus claim 9 wherein the active reciprocating fluid conveyance capsule is stacked and locked in any order and any number within the column and comprising liquid nutrient media and micro organisms, the liquid media further corresponding with one or more containers capable of receiving and despatching the liquid media in a such a way that the liquid media is conveyed in reciprocating directions through at least said top capsule and bottom capsule.
 11. The apparatus of claim 1 wherein the active fluid conveyance capsule is a rotating wheel pump capsule.
 12. The apparatus of claim 11 wherein the rotating wheel pump capsule is stacked and locked in any order and any number within the column and comprising liquid nutrient media and micro organisms, the liquid media further corresponding with one or more containers capable of receiving and despatching the liquid media in a such a way that the liquid media is conveyed in reciprocating directions through at least said top capsule and bottom capsule.
 13. A purification apparatus having a top capsule and a bottom capsule, the apparatus comprising a processing capsule having a purifying device for purifying a biologic product, and a second downstream capsule, wherein said capsules are stacked and locked in a column, with the proviso that the top capsule or bottom capsule are not pump capsules.
 14. The apparatus of claim 13 wherein the purifying device is in the top capsule.
 15. The apparatus of claim 13 further comprising an external fluid conveyance device, such as a pump means, connected to the processing capsule.
 16. The apparatus of claim 13 comprising liquid media and the biologic product.
 17. The apparatus of claim 13 wherein the second down-stream capsule is selected from a separation capsule, filtration capsule, chromatography capsule, ion exchange capsule, conditioning capsule, sparging capsule, diffusion capsule, active fluid conveyance capsule, passive fluid conveyance capsule, fluid collecting container, valve capsule, column end capsule, manifold capsule, storage capsule, power supply, data acquisition and control capsule.
 18. The apparatus of claim 13, comprising a further down-stream function selected from separation, filtration, chromatography, ion exchange, conditioning, sparging, diffusion, active fluid conveyance, passive fluid conveyance, valve, column end, manifold, storage functions, heating, cooling, power supply, data acquisition and controlling wherein the down-stream function or functions are located in one or more capsules.
 19. A bio factory apparatus assembly for producing a purified biologic product comprising a bioreactor apparatus and a purification apparatus in one column having a top capsule and a bottom capsule, the apparatus comprising a cultivation capsule having a production device for producing a biologic product, and a processing capsule having a purifying device for purifying a biologic product, wherein said capsules are stacked and locked in the column, with the proviso that the top capsule and bottom capsule are not pump capsules at the same time
 20. (canceled)
 21. The apparatus according to claim 19, wherein at least two of the capsules are separated by a sealing arrangement creating a boundary between said two capsules creating an internal sterile environment further separating the two capsules internal functional devices against the external non sterile environment.
 22. The apparatus according to claim 21, wherein the sealing arrangement is a gasket.
 23. The apparatus according to claim 22, wherein the gasket is prepared from an elastic organic material, such as rubber or a polymeric material with hardness ranging from 20-90 shore A.
 24. The apparatus according to claim 23, wherein the gasket is an inflatable gasket containing a closed volume to receive through a connection a pressurised fluid for expansion of the flexible gasket.
 25. The apparatus according to claim 19, wherein the biologic product is selected from Anti-coagulants, Blood factors, Cytokines, Fusion proteins, Growth factors, Hormones, Monoclonal antibodies, Polysaccharide vaccines, Recombinant vaccines, Therapeutic enzymes, Therapeutic antibodies, and Stem cells
 26. The apparatus of claim 19 wherein an up-stream or down-stream function is provided in a capsule pre-packed in a film bag.
 27. The apparatus of claim 19 wherein a capsule or capsule cover or parts of the capsule is pre-sterilized before use.
 28. The apparatus of claim 19 wherein a capsule or capsule cover or parts of the capsule is for single-use.
 29. The apparatus of claim 19 wherein a capsule or capsule cover or parts of the capsule is re-usable.
 30. A method of preparing a biologic product comprising the steps of: Cultivating a micro organism in a bioreactor apparatus of claim 1 and collecting the biological product.
 31. A method of purifying a biologic product comprising the steps of: Applying the un-purified biological product to the purifying apparatus of claim 13, and collecting the biological product in purified form.
 32. A method of producing a purified biologic product comprising the steps of: Cultivating a micro organism in a bio factory apparatus assembly claim 19, and collecting the biological product in purified form.
 33. A purification apparatus having a first capsule cover part comprising a first chamber having a conduit, a second capsule cover part comprising a second chamber having a conduit and one or more purification cartridges, wherein the first and second cover parts comprises one or more sets of couplings for axial integration of the purification cartridges, and wherein the conduits may be closed or in fluid connection with a capsule of a bioreactor apparatus or a further purification apparatus.
 34. The apparatus according to claim 33, wherein the conduit of the first capsule cover part or the conduit of the second capsule cover part has a sealing arrangement creating a boundary to the capsule of the bioreactor apparatus or purification apparatus, thereby creating an internal sterile environment further separating the internal functional devices against the external non sterile environment.
 35. The apparatus according to claim 33 wherein the couplings are tri-clamp ports or dual purpose cylindrical push-in first female function tube ports or single purpose cylindrical push-in tube ports or hose connections.
 36. The apparatus according to claim 33 wherein the first and second chamber is flat and circular.
 37. The apparatus according claim 33 wherein the first and second cover parts are identical.
 38. The apparatus according to claim 33 wherein the first and second cover parts are non-identical.
 39. The apparatus according to claim 33 wherein the purification cartridges are selected from commercially available cartridges, such as size exclusion cartridges or separation cartridges or chromatography capture cartridge or ion exchange cartridges.
 40. The apparatus according to claim 33 wherein the first or second chamber is arranged for fluid correspondence via the conduit to a down-stream or up-stream capsule. 41-43. (canceled)
 44. A method of preparing a biologic product comprising the steps of: Cultivating a micro organism in a bioreactor apparatus of claim 19 and collecting the biological product. 